用于检测基质金属蛋白酶的酶谱技术

Zymographical techniques for detection of matrix metalloproteinases.

作者信息

Iwai Shinichi, Nakanishi-Ueda Takako, Armstrong Donald, Oguchi Katsuji

机构信息

Department of Biotechnology and Clinical Laboratory Science, University at Buffalo, Buffalo, NY, USA.

出版信息

Methods Mol Biol. 2008;477:113-21. doi: 10.1007/978-1-60327-517-0_10.

DOI:10.1007/978-1-60327-517-0_10

PMID:19082943

Abstract

Matrix metalloproteinases (MMPs) are a family of zinc-dependent proteinases associated with extracellular matrix degradation, cellular migration, tissue remodeling, and angiogenesis. The activity of MMPs is regulated by the tissue inhibitors of metalloproteinases (TIMPs). Zymography and reverse zymography are useful to detect MMPs and TIMPs activities from various samples, for example vitreous, retina, plasma, and so on. Sample proteins are separated in substrate containing polyacrylamide gel by electrophoresis. The gel is incubated and then stained with Coomassie Blue. MMPs' activities are detected as clear bands.

摘要

基质金属蛋白酶（MMPs）是一类与细胞外基质降解、细胞迁移、组织重塑和血管生成相关的锌依赖性蛋白酶。MMPs的活性受金属蛋白酶组织抑制剂（TIMPs）的调节。酶谱法和反向酶谱法可用于检测来自各种样品（如玻璃体、视网膜、血浆等）的MMPs和TIMPs活性。样品蛋白通过电泳在含有聚丙烯酰胺凝胶的底物中进行分离。将凝胶孵育，然后用考马斯亮蓝染色。MMPs的活性以清晰的条带形式检测出来。